DNA methylation patterns in human iPSC-derived sensory neuronal differentiation.
Sensory neurons of the peripheral nervous system are critical in health and disease. Sensory neurons derived from induced pluripotent stem (iPS) cells are now being used increasingly for in vitro models of neuropathy, pain, and neurotoxicity. DNA methylation is critical for neurodevelopment and has been implicated in many neuronal diseases, but has not been examined in iPS-derived sensory neurons. In order to better characterize the iPS-derived sensory neuron model, we have undertaken a genome-wide DNA methylation study on the cells from human iPS to iPS-derived sensory neurons during differentiation through reduced representation and bisulfite sequencing. We report decreasing DNA methylation with iPS-derived sensory neuronal differentiation that is reflected in increasing numbers and proportions of hypomethylated individual CpGs and regions, as well as lowered DNMT3b expression. Furthermore, genes with changes in DNA methylation near their TSS suggest key pathways that may be involved in iPS-derived sensory neuronal differentiation. These findings provide insights into sensory neuronal differentiation and can be used for further in vitro modelling of disease states.


Morphine activates blast-phase chronic myeloid leukemia cells and alleviates the effects of tyrosine kinase inhibitors.
Morphine is an opioid analgesic drug routinely used in the postoperative period for pain management in cancer patients. In this work, we analyzed the effects of morphine on leukemia cells at all stages of development and addressed its underlying mechanism. We showed that clinically relevant concentrations of morphine promoted growth without affecting survival in blast phase-chronic myeloid leukemia (BP-CML) K562 and LAMA84 cells. In addition, morphine alleviated the anti-proliferative and pro-apoptotic effects of BCR-ABL tyrosine kinase inhibitor (TKI) in BP-CML cells. We further found that morphine increased colony formation and replating capacity of CD34 stem/progenitor derived from BP-CML patients. In addition, morphine alleviated the inhibitory effects of BCR-ABL TKIs in cell survival, colony formation and replating capacity in BP-CML CD34 + stem/progenitor cells. Mechanistic investigations demonstrated that morphine specifically activated Wnt signaling via increasing β-catenin activity and Wnt target genes transcription in K562 and CD34 + stem/progenitor cells. The effects of morphine in BP-CML were abolished by Wnt inhibitor LGK-974 or XAV939, which further confirmed that morphine protected BP-CML cells from BCR-ABL TKIs-induced toxicity via activating Wnt/β-catenin signaling. Our work demonstrated the novel role of morphine on leukemia cells. The activation of Wnt/β-catenin by morphine may provide a new guide in the clinical use of morphine, particularly in patients with Wnt-driven cancers.


Violations of safe diving practices among 122 diver fatalities.
Diving is a popular recreation with an excellent safety record, with an estimated 1.8 deaths per 1 million dives. This study investigated the relationship between intentional deviation from accepted diving practices (violations) and diver fatalities. The authors examined 119 incidents/122 diver fatalities that did not involve diver training in North America and the Caribbean, and identified the presence of violations of accepted diving safety practices, as well as if the death was associated with an acute medical event such as heart attack. Of the 122 fatalities, 57% (n = 70) were associated with a medical event and 43% (n = 52) were non-medical. Violations were found in 45% of fatalities (n = 55) overall. Violations were recorded for 23% of the 70 medical and 75% of the 52 non-medical fatalities. Divers who died from something other than a medical cause were 7 times as likely to have one or more violations associated with the fatality (OR 7.3, 95% CI 2.3-23.2). The odds of dying from something other than a medical condition increased approximately 60% for each additional 10 metres of depth. The odds of a death being associated with a medical condition increased approximately 9% per year of age, or 2.4 times for every 10 years older a diver was. Medical events are associated with over half of the non-training related diver fatalities in North America and the Caribbean, with the odds of death being associated with a medical condition doubling each decade of additional age. These data support recommendations that divers stay physically fit and have regular medical checkups, particularly as they get older. They also strongly support the safety benefit of adhering to established safe diving practices.


Estrogen biphasically modifies hypothalamic GABAergic function concomitantly with negative and positive control of luteinizing hormone release.
The principal role of estrogen is its control of the female ovulatory cycle via negative and positive feedback on gonadotropin secretion. However, a detailed, cohesive picture of how the steroid specifically regulates the excitability of hypothalamic neurons involved in the central control of gonadotropin secretion is still emerging. Here, we used an ovariectomized female guinea pig model to test the hypothesis that estrogen acts on GABAergic neurons in the preoptic area (POA) to elicit a biphasic profile of luteinizing hormone (LH) secretion. Intracellular electrophysiological recordings revealed that estradiol benzoate (EB; 25 microgram, s.c.) decreased the hyperpolarizing response of GABAergic neurons to the GABA(B) receptor agonist baclofen 24 hr after treatment. This effect of GABA(B) receptor stimulation in unidentified POA neurons was still depressed 42 hr after EB administration. By the use of a ribonuclease protection assay, however, EB reduced glutamic acid decarboxylase mRNA expression 42 hr but not 24 hr after its administration. Thus, estrogen attenuated the autoinhibition of GABAergic POA neurons during the initial LH suppressive (i.e., negative feedback) phase and subsequently reduced GABAergic function during the LH surge (i.e., positive feedback). These studies demonstrate that the effects of estrogen on hypothalamic GABAergic neurons coincide with the inhibitory and stimulatory actions, respectively, of the steroid on LH secretion. Furthermore, the data provide novel insights into the mechanism by which estrogen regulates hypothalamic GABAergic neurons, which are critical for the biphasic modulation of LH release observed over the course of the female ovulatory cycle.


Purification and identification of sperm surface proteins and changes during epididymal maturation.
Surface membrane proteins have a key role in the sequential interactions between spermatozoa and oocytes. The aim of this study was to characterize protein changes occurring during post-testicular differentiation using a new overall approach to study surface membrane proteins of spermatozoa. A dedicated protocol based on specific purification of surface membrane proteins labeled with sulfo-NHS-SS-biotin was developed for this purpose. Appropriate gel electrophoresis separation and purification methods combined with standard proteomic methods were then used to identify and quantify surface membrane proteins from immature and mature spermatozoa. Membrane-associated proteins were discriminated from integral membrane proteins by differential solubilization. Protein regionalization on the spermatozoon surface was achieved by comparative analysis of the surface protein extracts from the entire spermatozoa and from periacrosomal sperm plasma membranes. Identification of several known proteins and of new proteins related to the process of epididymal maturation showed the reliability of this protocol for specific purification of a subproteome and identification of new sperm membrane proteins. This approach opens up a new area in the search for male fertility markers.


Structure-based mechanism of preferential complex formation by apoptosis signal-regulating kinases.
Apoptosis signal-regulating kinases (ASK1, ASK2, and ASK3) are activators of the p38 and c-Jun N-terminal kinase (JNK) mitogen-activated protein kinase (MAPK) pathways. ASK1-3 form oligomeric complexes known as ASK signalosomes that initiate signaling cascades in response to diverse stress stimuli. Here, we demonstrated that oligomerization of ASK proteins is driven by previously uncharacterized sterile-alpha motif (SAM) domains that reside at the carboxy-terminus of each ASK protein. SAM domains from ASK1-3 exhibited distinct behaviors, with the SAM domain of ASK1 forming unstable oligomers, that of ASK2 remaining predominantly monomeric, and that of ASK3 forming a stable oligomer even at a low concentration. In contrast to their behavior in isolation, the ASK1 and ASK2 SAM domains preferentially formed a stable heterocomplex. The crystal structure of the ASK3 SAM domain, small-angle x-ray scattering, and mutagenesis suggested that ASK3 oligomers and ASK1-ASK2 complexes formed discrete, quasi-helical rings through interactions between the mid-loop of one molecule and the end helix of another molecule. Preferential ASK1-ASK2 binding was consistent with mass spectrometry showing that full-length ASK1 formed hetero-oligomeric complexes incorporating large amounts of ASK2. Accordingly, disrupting the association between SAM domains impaired ASK activity in the context of electrophilic stress induced by 4-hydroxy-2-nonenal (HNE). These findings provide a structural template for how ASK proteins assemble foci that drive inflammatory signaling and reinforce the notion that strategies to target ASK proteins should consider the concerted actions of multiple ASK family members.


The brightness-weight illusion.
Bigger objects look heavier than smaller but otherwise identical objects. When hefted as well as seen, however, bigger objects feel lighter (the size-weight illusion), confirming that the association between visual size and weight has a perceptual component. Darker objects also look heavier than brighter but otherwise identical objects. It is uncertain, however, if this association also has a perceptual element, or if it simply reflects the fact that, in English at least, the same verbal label (light) is applied to both surface brightness and weight. To address this, we looked for a brightness equivalent of the size-weight illusion. Paired-comparison judgments of weight were obtained for balls differing only in color. Based on vision alone, darker objects were judged to be heavier. When the balls were hefted as well as seen, this association was reversed (i.e., a brightness-weight illusion), consistent with it having a perceptual component. To gauge the strength of the illusion (in grams), a white and a black ball (both 129 g) were each compared against a set of mid-gray balls varying in weight. When the balls were hefted as well as seen, the white ball felt approximately 8 g heavier than the black ball, a difference corresponding to 6.2% of their actual weight. Possible environmental origins of the association between surface lightness and weight are considered.


Exercise-associated thermographic changes in young and elderly subjects.
This study aimed at evaluating the thermographic changes associated with localized exercise in young and elderly subjects. An exercise protocol using 1 kg load was applied during 3 min to the knee flexors of 14 elderly (67 +/- 5 years) and 15 young (23 +/- 2 years) healthy subjects. The posterior thigh's skin temperature of the exercised limb and contralateral limb were measured by infrared thermography on pre-exercise, immediately post-exercise, and during the 10-min period post-exercise. Difference (p < 0.01) between elderly and young subjects was observed on pre-exercise temperature. Although differences were not observed between pre-exercise and immediately post-exercise temperature in the exercised limb, thermographic profile displayed heat concentration in exercised areas for both groups. Temperature reduction was only observed for the young group on the 10-min post-exercise (p < 0.05) in the exercised limb (30.7 +/- 1.7 to 30.3 +/- 1.5 degrees C). In contrast, there was a temperature reduction post-exercise (p < 0.01) in the contralateral limb for both groups. These results present new evidences that elderly and young subjects display similar capacity of heat production; however, the elderly subjects presented a lower resting temperature and slower heat dissipation. This work contributes to improve the understanding about temperature changes in elderly subjects and may present implications to the sports and rehabilitation programs.


Female sexual function and gestational diabetes.
The number of women with gestational diabetes mellitus (GDM) is growing worldwide in parallel with the obesity epidemic. The diagnosis of GDM leads to substantial modifications in the daily routine of these women, and these adjustments could potentially affect their sexual function. There are no previous studies on the sexual function of patients with GDM. The aim of this study was to investigate the sexual function of patients with GDM in comparison with healthy pregnant women at the same gestational age. Brazilian women in the third trimester of pregnancy with and without GDM were invited to participate in this cross-sectional study while waiting for their antenatal care visits at a single public tertiary teaching institution between March and December 2010. The Brazilian version of the Female Sexual Function Index (FSFI) questionnaire was used to assess sexual function. Desire, arousal, lubrication, orgasm, sexual satisfaction, and pain during and after coitus in the last 4 weeks, measured according to a standardized and validated questionnaire. A total of 87 participants were enrolled (43 healthy women and 44 with GDM). There were no significant differences in the sociodemographic characteristics of both groups. The total FSFI scores of GDM patients was 21.0±9.59 compared with 22.3±9.17 for healthy women (P=0.523). Difficulty in desire was the most common sexual dysfunction symptom in both groups, being reported by 42% and 50% of GDM and healthy women, respectively (P=0.585). The sexual function of Brazilian patients with GDM does not differ significantly from that of healthy pregnant women at the same gestational age.


Stress tolerant virulent strains of Cronobacter sakazakii from food.
Cronobacter sakazakii is considered as an emerging foodborne pathogen. The aim of this study was to isolate and characterize virulent strains of Cronobacter sakazakii from food samples of Bangladesh. Six (6) Cronobacter sakazakii was isolated and identified from 54 food samples on the basis of biochemical characteristics, sugar fermentation, SDS-PAGE of whole cell protein, plasmid profile and PCR of Cronobacter spp. specific genes (esak, gluA, zpx, ompA, ERIC, BOX-AIR) and sequencing. These strains were found to have moderately high antibiotic resistance against common antibiotics and some are ESBL producer. Most of the C. sakazakii isolates were capable of producing biofilm (strong biofilm producer), extracellular protease and siderophores, curli expression, haemolysin, haemagglutinin, mannose resistant haemagglutinin, had high cell surface hydrophobicity, significant resistance to human serum, can tolerate high concentration of salt, bile and DNase production. Most of them produced enterotoxins of different molecular weight. The isolates pose significant serological cross-reactivity with other gram negative pathogens such as serotypes of Salmonella spp., Shigella boydii, Shigella sonnei, Shigella flexneri and Vibrio cholerae. They had significant tolerance to high temperature, low pH, dryness and osmotic stress. Special attention should be given in ensuring hygiene in production and post-processing to prevent contamination of food with such stress-tolerant virulent Cronobacter sakazakii.


Contribution of Gardnerella vaginalis to vaginitis in a general practice.
In a study of 154 adult women who presented to their general practitioner with vaginal symptoms 30 (20%) had Gardnerella vaginalis on its own and 51 (33%) had G vaginalis in combination with anaerobes or known pathogens. Thirty one (20%) patients were culture negative. Those who were culture negative had fewer symptoms and signs of vaginitis than those with G vaginalis alone or G vaginalis plus anaerobes. Those with known pathogens had more symptoms and signs than those with G vaginalis alone or G vaginalis plus anaerobes. Those with known pathogens plus G vaginalis had the most severe signs and symptoms of vaginitis. It is concluded that G vaginalis can cause vaginitis on its own, and it makes vaginitis worse when present with other organisms. G vaginalis was also found in 30 (21%) of the 138 control patients who, although they presented "asymptomatically," had worse signs than control patients without G vaginalis. It seems that G vaginalis can occur in a spectrum ranging from the uncomplaining patient to those with severe vaginitis.


Application of thrombin based fibrin glue and non-thrombin based batroxobin glue on intact human blood vessels: evidence for transmural thrombin activity.
An alternative method of uniting small diameter vessels to obtain tissue union while limiting the thrombogenic effect of suture placement at a vessel anastomosis involves the use of a thrombin based fibrin glue as a surgical sealant. This investigation addresses whether the in vitro application of a thrombin based glue (TG), or batroxobin glue (BG), a non-thrombin based glue made with the snake venom enzyme batroxobin, alters intravascular platelet deposition (PD) or cleaves blood fibrinogen, as measured by fibrinopeptide A (FPA) production, when the respective glue is applied to the external surface of an intact human placental artery or an artery with an anastomosis. When TG was applied to the adventitial surface of an intact vessel or an anastomosis (n = 7) of control and experimental vessels, there was a significant increase in intraluminal platelet deposition, an effect not realized with BG (n = 12, intact vessel TG p = 0.01, BG p = 0.66, anastomosis TG p <0.01, BG p <0.01). Both TG and BG significantly increased FPA levels when human whole blood was perfused through both intact vessels or vessels containing an anastomosis when compared to control vessels (intact vessel TG and BG p <0.01, anastomosis TG and BG p <0.01). Labelled thrombin studies document the rapid passage of thrombin through an intact vessel wall or vessels with an anastomosis when TG was applied to the adventitial surface of the vessel. The data suggest that TG and BG are drug delivery systems for their respective enzymes that either pass through or transfer a message across not only a surgically created anastomosis, but also an intact vessel wall.


Monolayered organosilicate toroids and related structures: A phase diagram for templating from block copolymers.
Here we report the controlled generation of micelle-templated organosilicate nanostructures resulting from self-assembly of a block copolymer/organosilicate mixture followed by organosilicate vitrification and copolymer thermolysis. Variation of solution condition and the copolymer/organosilicate mixture composition generates widely different film morphologies ranging from toroids to linear features to contiguous nanoporous monolayers. The use of reactive organosilicates for block copolymer templation generates functional inorganic nanostructures with thermal and mechanical stability.


Minds, Brains, and Capacities: Situated Cognition and Neo-Aristotelianism.
This article compares situated cognition to contemporary Neo-Aristotelian approaches to the mind. The article distinguishes two components in this paradigm: an Aristotelian essentialism which is alien to situated cognition and a Wittgensteinian "capacity approach" to the mind which is not just congenial to it but provides important conceptual and argumentative resources in defending social cognition against orthodox cognitive (neuro-)science. It focuses on a central tenet of that orthodoxy. According to what I call "encephalocentrism," cognition is primarily or even exclusively a computational process occurring inside the brain. Neo-Aristotelians accuse this claim of committing a "homuncular" (Kenny) or "mereological fallacy" (Bennett and Hacker). The article explains why the label "fallacy" is misleading, reconstructs the argument to the effect that encephalocentric applications of psychological predicates to the brain and its parts amount to a category mistake, and defends this argument against objections by Dennett, Searle, and Figdor. At the same time it criticizes the Neo-Aristotelian denial that the brain is the organ of cognition. It ends by suggesting ways in which the capacity approach and situated cognition might be combined to provide a realistic and ecologically sound picture of cognition as a suite of powers that flesh-and-blood animals exercise within their physical and social environments.


Implementation of treatment escalation plans in an old age psychiatry inpatient hospital.
A treatment escalation plan (TEP) enables timely and appropriate decision making in the management of deteriorating patients. The COVID-19 pandemic precipitated the widespread use of TEPs in acute care settings throughout the National Health Service (NHS) to facilitate safe and effective decision making. TEP proformas have not been developed for the inpatient psychiatric setting. This is particularly concerning in old age psychiatry inpatient wards where patients often have multiple compounding comorbidities and complex decisions regarding capacity are often made. Our aim for this quality improvement project was to pilot a novel TEP proforma within a UK old age psychiatry inpatient hospital. We first adapted a TEP proforma used in our partner acute tertiary hospital and implemented it on our old age psychiatry wards. We then further refined the form and gathered data about uptake, length of time to complete a TEP and the ceiling of care documented in the TEP. We also explored staff, patient and family views on the usefulness of TEP proformas using questionaries. TEP decisions were documented in 54% of patient records at baseline. Following revision and implementation of a TEP proforma this increased to 100% on our two wards. The mean time taken to complete a TEP was reduced from 7.1 days to 3.2 days following inclusion of the TEP proforma in admission packs. Feedback from staff showed improvements in understanding about TEP and improved knowledge of where these decisions were documented. We advocate the use of TEP proformas on all old age psychiatry inpatient wards to offer clear guidance to relatives and treating clinicians about the ceilings of care for patients. There are potentially wider benefits to healthcare systems by reducing inappropriate transfers between psychiatry and acute NHS hospitals.


Cost-benefit analysis of a strategy to vaccinate healthy working adults against influenza.
Influenza is a major cause of illness, disruption to daily life, and work absenteeism among healthy working adults aged between 18 and 64 years. This group is not included among the traditional priority groups for annual vaccination. Immunization rates remain low. To assess the economic implications of a strategy for annual vaccination of this group. Using the societal perspective, this cost-benefit analysis included the direct and indirect costs associated with vaccination as well as the direct and indirect costs prevented by vaccination. Clinical and economic variable estimates were derived primarily from the published literature. For this model, it was assumed that vaccination occurred in efficient, low-cost settings such as at the work site. Monte Carlo simulation was used to calculate the mean net costs or savings along with the 95% probability interval, and sensitivity analyses explored the sensitivity of the cost model to different values of the input variables. Vaccinating healthy working adults was on average cost saving, with mean savings of $13.66 per person vaccinated (95% probability interval: net savings of $32.97 to net costs of $2.18), with vaccination generating net savings 95% of the time. The model was most sensitive to the influenza illness rate, the work absenteeism rate due to influenza, and hourly wages. In the worst-case scenario vaccination was not cost saving. Vaccination also generated net costs to society during years with a poor vaccine-circulating virus strain match. In all of the other sensitivity analysis scenarios, vaccination was cost saving. Influenza vaccination of healthy working adults on average is cost saving. These findings support a strategy of routine, annual vaccination for this group, especially when vaccination occurs in efficient and low-cost sites.


Impact of minimum hematocrit during cardiopulmonary bypass on mortality in patients undergoing coronary artery surgery.
The hematocrit on cardiopulmonary bypass (CPB) frequently falls to a low level during many cardiac surgical procedures. This study was designed to explore the impact on mortality of minimum hematocrit level achieved during the CPB after coronary artery surgery. Two thousand seven hundred thirty-eight sequential isolated coronary artery surgery patients during a 42-month period at a tertiary academic center were included in this study. Thirty-one standardized preoperative risk factors used in a multiple logistic regression revealed eight statistically significant independent predictors for postoperative mortality. Minimum hematocrit level during CPB was then added to the regression model and was found to be an independent risk factor for mortality. The entire patient population was divided into dichotomous groups using different minimum hematocrit levels on CPB for the determination of cutoff points by multiple logistic regression. After adjusting for other risk factors, the minimum hematocrit level of 14% was found to be a statistically significant cutoff point. Patients with minimum hematocrit levels < or =14% were found to have an increased probability of risk-adjusted mortality (odds ratio, 2.70; P=.002). A subgroup analysis revealed that high-risk patients with minimum hematocrit levels < or =17% were found to have a significantly increased probability of postoperative mortality (odds ratio, 2.20; P=.017). Minimum hematocrit level during CPB is an independent risk factor for mortality after coronary artery surgery. There is a significantly increased risk of mortality for hematocrit levels < or =14%. For high-risk patients, there is a significantly increased risk of mortality for hematocrit levels < or =17%.


Mechanistic studies of the copolymerization reaction of oxetane and carbon dioxide to provide aliphatic polycarbonates catalyzed by (Salen)CrX complexes.
Chromium salen derivatives in the presence of anionic initiators have been shown to be very effective catalytic systems for the selective coupling of oxetane and carbon dioxide to provide the corresponding polycarbonate with a minimal amount of ether linkages. Optimization of the chromium(III) system was achieved utilizing a salen ligand with tert-butyl groups in the 3,5-positions of the phenolate rings and a cyclohexylene backbone for the diimine along with an azide ion initiator. The mechanism for the coupling reaction of oxetane and carbon dioxide has been studied. Based on binding studies done by infrared spectroscopy, X-ray crystallography, kinetic data, end group analysis done by (1)H NMR, and infrared spectroscopy, a mechanism of the copolymerization reaction is proposed. The formation of the copolymer is shown to proceed in part by way of the intermediacy of trimethylene carbonate, which was observed as a minor product of the coupling reaction, and by the direct enchainment of oxetane and CO 2. The parity of the determined free energies of activation for these two processes, namely 101.9 kJ x mol (-1) for ring-opening polymerization of trimethylene carbonate and 107.6 kJ x mol (-1) for copolymerization of oxetane and carbon dioxide supports this conclusion.


Mucus glycoproteins from cystic fibrotic sputum. Macromolecular properties and structural 'architecture'.
Mucus glycoproteins (mucins) were isolated from sputum of patients with cystic fibrosis (CF) after separation into sol and gel phases. The mucus gel was solubilized with gentle stirring in 6 M-guanidinium chloride supplemented with proteinase inhibitors, and purification of mucins was subsequently achieved by isopycnic density-gradient centrifugation in CsCl/guanidinium chloride. Density-gradient centrifugation also revealed a heterogeneity of the macromolecules, the pattern of which varied between individuals, and mucins from the gel phase was pooled as 'heavy' and 'light' fractions. Gel chromatography on Sepharose CL-2B showed that the heavy fraction contained a larger proportion of smaller species than the 'light' fraction and that the gel phase mucins were much larger than those from the sol. An apparently homogeneous high-Mr mucin population from one individual contained approx. 70% (w/w) carbohydrate, the major sugars being N-acetylglucosamine (17.8%), N-acetylgalactosamine (6.7%), galactose (20.7%), fucose (13.2%) and sialic acid (11.4%). These mucins had an S020.w of 47 S, and an Mr of 15 x 10(6) -20 x 10(6), and rate-zonal centrifugation revealed a polydisperse size distribution [range (5-30) x 10(6)] with a weight-average Mr of 17 x 10(6). The whole mucins were visualized with electron microscopy as linear and apparently flexible threads, disperse in size. Reduction produced subunits which were included on Sepharose CL-2B, and subsequent trypsin digestion yielded high-Mr glycopeptides which were further retarded. The size distributions and fragmentation patterns of mucin from two other CF patients were the same, as studied by gel chromatography, rate-zonal centrifugation and electron microscopy. We conclude that CF mucins are heterogeneous in both size and buoyant density and that the various populations, though differing in buoyant density, share the same architecture and macromolecular properties and are, in this respect, similar to mucins from normal respiratory secretions [Thornton, Davies, Kraayenbrink, Richardson, Sheehan & Carlstedt (1990) Biochem. J. 265, 179-186] and human cervical mucus [Carlstedt & Sheehan (1989) SEB Symp. XLIII 289-316].


The Dominant white, Dun and Smoky color variants in chicken are associated with insertion/deletion polymorphisms in the PMEL17 gene.
Dominant white, Dun, and Smoky are alleles at the Dominant white locus, which is one of the major loci affecting plumage color in the domestic chicken. Both Dominant white and Dun inhibit the expression of black eumelanin. Smoky arose in a White Leghorn homozygous for Dominant white and partially restores pigmentation. PMEL17 encodes a melanocyte-specific protein and was identified as a positional candidate gene due to its role in the development of eumelanosomes. Linkage analysis of PMEL17 and Dominant white using a red jungle fowl/White Leghorn intercross revealed no recombination between these loci. Sequence analysis showed that the Dominant white allele was exclusively associated with a 9-bp insertion in exon 10, leading to an insertion of three amino acids in the PMEL17 transmembrane region. Similarly, a deletion of five amino acids in the transmembrane region occurs in the protein encoded by Dun. The Smoky allele shared the 9-bp insertion in exon 10 with Dominant white, as expected from its origin, but also had a deletion of 12 nucleotides in exon 6, eliminating four amino acids from the mature protein. These mutations are, together with the recessive silver mutation in the mouse, the only PMEL17 mutations with phenotypic effects that have been described so far in any species.


Feasibility of eyes open alpha power training for mental enhancement in elite gymnasts.
This study focuses on a novel, easy to use and instruction-less method for mental training in athletes. Previous findings suggest that particular mental capacities are needed for achieving peak performance; including attentional control, focus, relaxation and positive affect. Electroencephalography (EEG) alpha brain activity has been associated with neural inhibition during processes of selective attention, for improving efficiency in information processing. Here we hypothesised that eyes open alpha power training by music teaches athletes to (1) learn to self-regulate their brain activity, and (2) learn to increase their baseline alpha power, herewith improving mental capacities such as focusing the allocation of attention. The study was double-blind and placebo-controlled. Twelve elite gymnasts were either given eyes open alpha power training or random beta power training (controls). Results indicate small improvements in sleep quality, mental and physical shape. In our first attempt at getting a grip on mental capacities in athletes, we think this novel training method can be promising. Because gymnastics is one of the most mentally demanding sports, we value even small benefits for the athlete and consider them indicative for future research.


Changes in body composition in apparently healthy urban Indian women up to 3 years postpartum.
Dietary and life style practices differ in postpartum (PP) and nonpregnant Indian women. Effect of these practices on postpartum weight retention (PPWR) and development of cardio-metabolic risk (CMR) has been scarcely studied in urban women. Aims of this study were to (i) compare anthropometry, biochemical parameters and body composition up to 3 years PP (ii) effect of PPWR, dietary fat intake and physical activity on CMR factors. Cross-sectional, 300-fullterm, apparently healthy primi-parous women (28.6 ± 3.4 years) randomly selected. 128 women within 7-day of delivery (Group-A), 88 with 1-2 years (Group-B) and 84 with 3-4-year-old-children (Group-C) were studied. Anthropometry, sociodemographic status, physical activity, diet, clinical examination, biochemical tests, body composition, at total body (TB), by dual energy X-ray absorptiometry (GE-Lunar DPX) were collected. Women at 3-year PP showed higher weight retention (6.5[10] kg) than at 1-year (3.0[7] kg) (median [IQR]). Android fat % (central obesity) increased (P < 0.05) at 1-year PP (47 ± 10.0%) when compared to 1-week PP (44.3 ± 6.7%) and remained elevated at 3-year PP (45.6 ± 10.2%). Regression analysis revealed that at 1-year PP, increase in PPWR (Odd Ratio [OR] 1.8, 95% confidence interval [CI] = [1.2, 2.5], P < 0.001) and inactivity (OR 1.4, 95% CI= (0.97, 2.0), P < 0.1) were predictors for CMR. At 3-year PP, only PPWR was responsible for increase in CMR parameters (OR 1.6, 95% CI = (1.3, 2.3), P < 0.001) and not inactivity (P > 0.1). Postdelivery, low physical activity and higher PPWR may increase CMR in Indian women.


Ethnic Identity in Diverse Schools: Preadolescents' Private Regard and Introjection in relation to Classroom Norms and Composition.
Ethnic identity plays a key role in the normative development of children and adolescents, and efforts to provide a positive and safe environment for ethnic identity benefit from an understanding of its context-dependency. Following the social identity perspective, we add to research on ethnic identity by considering the role of the classroom context and by conceptualizing ethnic identity in terms of two key dimensions. Specifically, the present study aims to investigate the role of the classroom context for ethnic private regard (positive ethnic self-feelings) and for the under-researched construct of ethnic introjection (subjective self-group merging). These two dimensions of ethnic identity were examined in 51 Dutch school classes among grade 4-6 students (N= 573; Mage = 10.77, SD = 1.02; 54% girls) of Dutch, Turkish and Moroccan ethnic background. We focused on teachers' multicultural norms and classmates' evaluation of the ethnic in-group (peer group norms) in combination with the ethnic class composition. It was found that ethnic introjection was empirically distinct from ethnic private regard, and that the former dimension depended on the classroom context more than the latter. Multicultural teacher norms affected minority preadolescents' private regard positively, but only when the share of in-group classmates was low. Positive peer group norms of in-group classmates strengthened students' introjection, while those of out-group classmates lowered it. The findings indicate that ethnic identity research will be enhanced by more fully considering the conceptual and contextual implications of the social identity perspective.


Development of a total colectomy and ileorectal anastomosis rat model to evaluate colonic metaplasia.
Ulcerative colitis is an idiopathic inflammatory condition of the colon that may require surgical intervention including proctocolectomy and either ileal pouch-anal anastomosis or in the pediatric population, low ileorectal anastomosis (IRA). Often, subsequent physiologic alteration (or colonic metaplasia) occurs in the anastomosed small bowel that includes changes in mucin content, villous blunting, and increased expression of WNT5A, a marker of colonic crypt regeneration. We developed a rat low IRA model to assess and study the development of colonic metaplasia. We subjected male Sprague-Dawley rats (n = 17) to total colectomy and low IRA surgery and evaluated healing periodically by endoscopic evaluation. The ileum upstream of the anastomosis was assessed by hematoxylin and eosin staining, and the mucin content was measured by high iron diamine-Alcian blue staining. Wnt5a transcripts were quantified by reverse transcription and quantitative polymerase chain reaction at the 8-wk study end point. Although no gross endoscopic evidence of inflammation was seen throughout the course of the study, colonic metaplasia in the small bowel was detected in 7 out of 10 (70%) rats at the study end point. In rats with colonic metaplasia, enhanced expression of Wnt5a was evident at the study end point compared to levels in the terminal ileum at the time of surgery. Within 4-8 wk, the majority of rats subjected to IRA developed colonic metaplasia defined by villous blunting, changes in mucin content, and increased expression of Wnt5a. This model provides a method to study small bowel colonic metaplasia.


Transient receptor potential (TRP) protein 7 acts as a G protein-activated Ca2+ channel mediating angiotensin II-induced myocardial apoptosis.
Transient receptor potential (TRP) proteins have been identified as cation channels that are activated by agonist-receptor coupling and mediate various cellular functions. TRPC7, a homologue of TRP channels, has been shown to act as a Ca2+ channel activated by G protein-coupled stimulation and to be abundantly expressed in the heart with an as-yet-unknown function. We studied the role of TRPC7 in G protein-activated signaling in HEK293 cells and cultured cardiomyocytes in vitro transfected with FLAG-tagged TRPC7 cDNA and in Dahl salt-sensitive rats with heart failure in vivo. TRPC7-transfected HEK293 cells showed an augmentation of carbachol-induced intracellular Ca2+ transient, which was attenuated under a Ca2+-free condition or in the presence of SK&F96365 (a Ca2+-permeable channel blocker). Upon stimulation with angiotensin II (Ang II), cultured neonatal rat cardiomyocytes transfected with TRPC7 exhibited a significant increase in apoptosis detected by TUNEL staining, accompanied with a decrease in the expression of atrial natriuretic factor and destruction of actin fibers, as compared with non-transfected cardiomyocytes. Ang II-induced apoptosis was inhibited by CV-11974 (Candesartan; Ang II type 1 [AT1] receptor blocker), SK&F96365, and FK506 (calcineurin inhibitor). In Dahl salt-sensitive rats, apoptosis and TRPC7 expression were increased in the failing myocardium, and a long-term treatment with temocapril, an angiotensin-converting enzyme inhibitor, suppressed both. Our findings suggest that TRPC7 could act as a Ca2+ channel activated by AT1 receptors, leading to myocardial apoptosis possibly via a calcineurin-dependent pathway. TRPC7 might be a key initiator linking AT1-activation to myocardial apoptosis, and thereby contributing to the process of heart failure.


Ultra-rapid delayed rectifier channels: molecular basis and therapeutic implications.
The ultrarapid delayed rectifier channels have attracted considerable interest as targets for 'atrial-selective' antiarrhythmic drugs because they contribute to atrial but not to ventricular repolarization. Thus, I(Kur) channel blockers are expected to prolong selectively the atrial effective refractory period without inducing proarrhythmic effects due to excessive ventricular action potential prolongation. Here we provide an overview of the properties of I(Kur) channels in expression systems and native cardiomyocytes. The ion conducting pore of the channel is formed by four Kv1.5 α-subunits, whereas the ancillary β-subunits Kvβ1.2, Kvβ1.3, and Kvβ2.1 control channel trafficking and plasma membrane integration as well as activation and inactivation kinetics. Investigation of I(Kur) channel blockers in cardiomyocytes is complicated (i) by substantial overlap of I(Kur) with other currents, notably the transient outward current I(to), (ii) by lack of drug selectivity, and (iii) by disease-induced regulation of I(Kur). Some new compounds developed as I(Kur) blockers are described and their efficacy in treatment of atrial fibrillation (AF) is discussed. Current evidence suggests that pure I(Kur) channel block may not be sufficient to suppress AF.


Biochemical changes in equine erythrocytes during experimental regenerative anemia.
Hemolytic or blood loss anemia was induce in six ponies and red blood cell concentrations of creatine, glucose-6-phosphate dehydrogenase (G-6-PD), lactate dehydrogenase (LDH), and aspartate transaminase (AST) were measured during the ensuing regenerative period. Creatine and G-6-PD levels correlated well and increased concentration of either was good indication of increased erythrogenesis. Erythrocyte LDH levels were of value in assessing the response to hemolytic anemia but not to blood loss anemia. The difference may be, at least in part, the result of differing degrees of regenerative effort seen in the two experimental groups. Red cell AST concentrations fluctuated markedly and were of no value in assessing the anemia in either group.


Aberrant production of IL-12 by macrophages from several autoimmune-prone mouse strains is characterized by intrinsic and unique patterns of NF-kappa B expression and binding to the IL-12 p40 promoter.
Intrinsic defects in macrophage (M(phi)) cytokine production characterize many autoimmune-prone mouse strains. Aberrant levels of IL-12, for example, are produced by M(phi) isolated from young mice prone to lupus (MRL and NZB/W) and diabetes (nonobese diabetic (NOD)) well before the appearance of disease signs. Evaluation of the possible mechanism(s) underlying the abnormal regulation of IL-12 in these strains revealed novel patterns of Rel family protein binding to the unique p40 NF-kappaB site in the IL-12 p40 promoter, whereas binding patterns to Ets and CCAAT enhancer binding protein/beta sites were normal. In particular, the heightened production of IL-12 by NOD M(phi) is associated with elevated levels of the trans-activating p50/c-Rel (p65) complex compared with the nonfunctional p50/p50 dimer. Conversely, the dramatically impaired production of IL-12 by both NZB/W and MRL/+ M(phi) is associated with a predominance of p50/p50 and reduced p50/c-Rel(p65) binding. Mechanistically, the unique pattern seen in the lupus strains reflects elevated p50 and reduced c-Rel nuclear protein levels. In NOD extracts, the level of c-Rel is elevated compared with that in lupus strains, but not when compared with that in normal A/J. However, the extent of c-Rel tyrosine phosphorylation noted in NOD extracts is more than double that seen in any other strain. Levels of p65 were similar in all strains tested. These findings reveal that a common mechanism, involving dysregulation of c-Rel and p50, may be used to determine the aberrant IL-12 levels that have the potential to predispose specific mouse strains to systemic or organ-specific autoimmunity.


Carotid baroreceptor unloading decreases plasma atrial natriuretic factor in hypertensive patients.
Atrial natriuretic factor (ANF) release is known to be regulated by distension of the atrial wall; other factors affecting ANF secretion have not yet been defined. In order to evaluate the effects of the reflex activation of the sympathetic nervous system on ANF plasma levels, in 11 patients with essential hypertension progressive deactivation of carotid baroreceptors was induced by 4-min graded increases in external neck tissue pressure. Carotid sinus hypotension induced progressive increases in blood pressure (BP), heart rate (HR) and forearm vascular resistance (FVR), while plasma renin activity and catecholamine concentrations did not change significantly. Despite the lack of changes in right atrial pressure during this manoeuvre, plasma ANF levels showed a progressive and significant reduction, which was correlated with the increase in FVR. Although a contribution by other factors cannot be ruled out, our data suggest that the reflex activation of sympathetic nervous system is associated with reduced ANF release, independent of changes in atrial pressure.


Activation Receptor-Dependent IFN-γ Production by NK Cells Is Controlled by Transcription, Translation, and the Proteasome.
NK cells can recognize target cells such as virus-infected and tumor cells through integration of activation and inhibitory receptors. Recognition by NK cells can lead to direct lysis of the target cell and production of the signature cytokine IFN-γ. However, it is unclear whether stimulation through activation receptors alone is sufficient for IFN-γ production. In this study, we show that NK activation receptor engagement requires additional signals for optimal IFN-γ production, which could be provided by IFN-β or IL-12. Stimulation of murine NK cells with soluble Abs directed against NK1.1, Ly49H, Ly49D, or NKp46 required additional stimulation with cytokines, indicating that a range of activation receptors with distinct adaptor molecules require additional stimulation for IFN-γ production. The requirement for multiple signals extends to stimulation with primary m157-transgenic target cells, which triggers the activation receptor Ly49H, suggesting that NK cells do require multiple signals for IFN-γ production in the context of target cell recognition. Using quantitative PCR and RNA flow cytometry, we found that cytokines, not activating ligands, act on NK cells to express Ifng transcripts. Ly49H engagement is required for IFN-γ translational initiation. Results using inhibitors suggest that the proteasome-ubiquitin-IKK-TPL2-MNK1 axis was required during activation receptor engagement. Thus, this study indicates that activation receptor-dependent IFN-γ production is regulated on the transcriptional and translational levels.


Personality strengths in romantic relationships: Measuring perceptions of benefits and costs and their impact on personal and relational well-being.
Three studies using samples of people in romantic relationships were conducted to create a new individual difference measure of partner strengths in couples. The 2 perceptions of partner strengths included (1) appreciation of their use and effectiveness and (2) recognition of costs associated with their use. Factor analyses supported 2-factors and we found that greater appreciation of partner strengths predicted greater relationship satisfaction, commitment, investment, intimacy, self-expansion, and support for goal pursuit; recognizing significant costs with partner strengths was inversely related to several outcomes. Using a 1-week daily diary, we found that appreciation of partner strength use and recognition of costs associated with these strengths predicted daily relationship satisfaction and whether basic psychological needs were met within the relationship. The explanatory power of partner strength perceptions could not be explained by the actual character strengths or Big Five personality traits of partners, support for positive self-disclosures (capitalization), or gratitude for relationship partners. Finally, we found that the relational consequences of partner strength perceptions were not just "in the head" of the perceiver-influencing partner relational outcomes. This research program provides evidence for the use of a new measure of how strengths are perceived to better understand romantic couples and aspirational targets in clinical interventions. (PsycINFO Database Record


Cholinergic neurons of the chick ciliary ganglia express adrenergic traits in vivo and in vitro.
In this study, we sought to determine whether neurons of the chick embryo ciliary ganglia (CG), a parasympathetic cholinergic ganglia, can express catecholaminergic (CA) traits. To accomplish this, we used immunocytochemical techniques to examine the presence of the CA enzymes tyrosine hydroxylase (TH) and phenylethanolamine N-methyltransferase (PNMT) in CGs removed from chick embryo at day 8 of development (E8). Few neurons containing TH but not PNMT were found in the E8 CG. To examine whether CG neurons express CA enzymes in vitro, CGs removed from E8 chick embryo were dissociated and kept in culture for 3 to 12 days. In 50% of the culture dishes, some neurons contain TH or PNMT after 5 days in vitro. In an equal proportion of culture plates, CG neurons did not express the enzymes. To determine whether the proportion of CG neurons expressing TH or PNMT is increased by tissue influences, ganglion cells were co-cultured with notochord. In 90% of the co-culture experiments, most neurons present in the culture dishes stained with TH or PNMT after 5 days in vitro. To test for the presence of aromatic L-amino acid decarboxylase (AADC), another CA enzyme, cultures of CGs and CGs plus notochord were incubated with levodopa and processed for the detection of CA histofluorescence. Dopamine histofluorescence was present in all neurons after 3 days in vitro irrespective of the presence of notochord, suggesting that the expressions of TH and PNMT and that of AADC are differentially regulated. This study, therefore, demonstrates that cholinergic neurons of the CG contain CA enzymes in vivo and in vitro and that the proportion of neurons expressing CA traits during development in vitro can be increased by environmental cues such as those released by the notochord.


Effects of Grade and School Services on Children's Responsibility for Hearing Aid Care.
Purpose We investigated trends in hearing aid maintenance and assumption of responsibility for hearing aids in school-age children who are hard of hearing. Specifically, we examined the extent to which families own necessary hearing aid maintenance equipment, whether and by whom hearing aid maintenance tasks are being completed, and the effects of grade and receipt of an Individualized Education Program (IEP) or 504 plan on a child's assumption of responsibility for hearing aid care. Method Participants included 167 children who are hard of hearing in 1st to 4th grade. Caregivers reported whether the families owned various types of hearing aid maintenance equipment (listening tube, battery tester, and dri-aid kit) and who normally completes various hearing aid maintenance tasks. Information about children's audiological characteristics was also collected. Results Thirty-two percent of families reported not owning at least 1 piece of hearing aid maintenance equipment. Using a battery tester and performing a listening check were the maintenance tasks completed the least frequently, with 49% and 28% of caregivers reporting that these tasks are not completed regularly, respectively. Children's responsibility for hearing aid maintenance increased with grade. After controlling for maternal education and degree of hearing loss, children with an IEP or 504 plan took more responsibility for hearing aid maintenance tasks than children without these services. Conclusion Important hearing aid maintenance tasks, such as listening checks, are not completed regularly for many children, even when families own the necessary equipment. Ensuring that children who are hard of hearing have an IEP or 504 plan throughout elementary school may support self-advocacy and encourage children to take responsibility for their hearing aids, which may lead to more consistent hearing aid functioning.


Persistent leukocytosis-is this a persistent problem for patients with acute ischemic stroke?
In the setting of acute ischemic stroke (AIS), leukocytosis has been shown to be an indicator of inflammatory response. Although leukocytosis on admission has been shown to correlate with initial stroke severity in AIS patients, no work has been done to assess if there are differences in transient or persistent leukocytosis in patients without infection. The objective of this study is to determine the clinical significance of persistent versus transient leukocytosis during the early phase of AIS. Patients who presented with AIS to our center within 48 hours of symptom onset between July 2008 and June 2010 were retrospectively identified by chart review. Patients were included if they had leukocytosis on admission (defined as white blood cell count >11,000/μL based on laboratory reference range values). A logistic regression model was used to evaluate persistent leukocytosis (leukocytosis 48 hours after admission) as a predictor of several outcome measures, including good functional outcome (discharge modified Rankin Scale score of 0-2). Marginal effects were used to estimate the probability of poor functional outcome. Of the 438 patients screened, 49 had leukocytosis on admission and of those 24 (49%) had persistent leukocytosis. NIHSS score correlated significantly with persistence of leukocytosis (r = .306; P = .0044). More people with transient leukocytosis (leukocytosis lasting <48 hours) had a good functional outcome (44% versus 16%; P = .006). After adjusting for baseline NIHSS score, persistent leukocytosis was not a significant independent predictor of good functional outcome, but showed an association (OR, 2.5; 95% CI, .562-10.7; P = .2322). Persistent leukocytosis after adjusting for age and NIHSS score at admission is associated with a poor functional outcome, but it is not statistically significant (OR, 2.43; 95% CI, .59-9.87; P = .2151). After controlling for age and NIHSS score on admission, for patients with persistent leukocytosis, the probability of having poor functional outcome at discharge was increased by 16 percentage points. Persistent leukocytosis is associated with higher baseline NIHSS scores. Persistent leukocytosis is tightly linked with baseline stroke severity and is associated with poor patient outcomes. Our study found that patients with persistent leukocytosis are more likely to present with severe strokes and maintain a high NIHSS score at 24 hours after admission, unlike patients without leukocytosis or patients with transient leukocytosis. Furthermore, it appears that persistent leukocytosis outside the setting of an infection negatively impacts the short-term functional outcome of AIS patients. Identifying patients with persistent leukocytosis could help to prognosticate and target patients that may benefit from future anti-inflammatory interventions.


Binding interactions between soluble HIV envelope glycoproteins and quaternary-structure-specific monoclonal antibodies PG9 and PG16.
PG9 and PG16 are antibodies isolated from a subject infected with HIV-1 and display broad anti-HIV neutralizing activities. They recognize overlapping epitopes, which are preferentially expressed on the membrane-anchored trimeric form of the HIV envelope glycoprotein (Env). PG9 and PG16 were reported not to bind to soluble mimetics of Env. The engineering of soluble Env proteins on which the PG9 and PG16 epitopes are optimally exposed will support efforts to elicit broad anti-HIV neutralizing antibodies by immunization. Here, we identified several soluble gp140 Env proteins that are recognized by PG9 and PG16, and we investigated the molecular details of those binding interactions. The IgG versions of PG9 and PG16 recognize the soluble trimeric gp140 form less efficiently than the corresponding monomeric gp140 form. In contrast, the Fab versions of PG9 and PG16 recognized the monomeric and trimeric gp140 forms with identical binding kinetics and with binding affinities similar to the high binding affinity of the anti-V3 antibody 447D to its epitope. Our data also indicate that, depending on the Env backbone, the interactions of PG9 and PG16 with gp140 may be facilitated by the presence of the gp41 ectodomain and are independent of the proper enzymatic cleavage of gp140 into gp120 and gp41. The identification of soluble Env proteins that express the PG9 and PG16 epitopes and the detailed characterization of the molecular interactions between these two antibodies and their ligands provide important and novel information that will assist in improving the engineering of future Env immunogens.


Esophageal polypoid dysplasia of gastric foveolar phenotype with focal intramucosal carcinoma associated with Barrett's esophagus.
We describe a rare case of esophageal polypoid dysplasia with gastric phenotype and focal intramucosal carcinoma associated with Barrett's esophagus. A 69-year-old man with a long history of gastroesophageal reflux disease was initially seen at an outside institution for evaluation of significant dysphagia. Screening upper gastrointestinal endoscopic evaluation revealed a large intraluminal polypoid lesion occluding the distal portion of the esophagus. Surgery was performed with resection of the distal esophagus and proximal stomach. The histopathologic examination of this lesion revealed an exuberant polypoid gastric epithelium with areas of low-grade dysplasia, high-grade dysplasia, and focal intramucosal carcinoma. A few residual foci of specialized intestinal metaplasia consistent with Barrett's esophagus without dysplasia were identified at the proximal and distal ends of the lesion. Immunohistochemically, this lesion revealed a pattern of expression of apomucins (MUC5AC diffusely positive, MUC1 and MUC6 focally positive, and MUC2 negative) consistent with a gastric foveolar phenotype. In addition, in the dysplastic areas, there was high Ki-67 labeling index and no overexpression of p53 protein. In our opinion, this case represents a precursor lesion of an extremely well-differentiated adenocarcinoma of gastric foveolar phenotype that has been previously documented in the stomach and in the duodenum and that now for the first time we report in the esophagus in association with Barrett's intestinal metaplasia.


A study of actinic cheilitis treatment by two low-morbidity CO2 laser vaporization one-pass protocols.
Therapeutic approaches to chronic actinic cheilitis focus on the removal or destruction of diseased epithelium. The CO(2) laser has become an important therapeutic alternative, achieving clinical resolution in around 90% of patients. Although many laser physical parameters have been reported, some are known for their low potential for scar induction without compromising the success of the results. The aim of this clinicohistological study was to compare the therapeutic responses to two low-morbidity protocols involving a single laser pass. A total of 40 patients with chronic multicentric and microscopically proven disease were randomly submitted to two conservative CO(2) laser protocols using a bilateral comparative model. The degree of histological atypia of the epithelium was determined in 26 patients both pre- and postoperatively for both protocols. Other histological phenomena were assessed in addition to this central analysis parameter. Clinical recurrence occurred in 12.5% of patients for each protocol, together with a significant reduction in the degree of epithelial atypia (p < 0.001), which was occasionally complete. However, no difference was found between the protocols (p > 0.05). Using these morphological parameters it was not possible to determine whether postoperative epithelial atypias in part of the sample were reactive or residual in nature. A few patients may show minor postoperative lesions. Due to their potential to achieve clinical and importantly microscopic resolution, the studied protocols may be used for mild through moderate dysplastic epithelium and clinically diffuse disease.


Spinal intradural cerebellar ectopia.
An ectopic cerebellum, as in Chiari malformations and ectopic cerebellar dysplastic tissue, is a common finding; however, the presence of an organized ectopic cerebellum is exceedingly rare. We describe the MR imaging, surgical, and histologic appearance of an intraspinal ectopic cerebellum in an infant.


Analysis of damage tolerance pathways in Saccharomyces cerevisiae: a requirement for Rev3 DNA polymerase in translesion synthesis.
The replication of double-stranded plasmids containing a single N-2-acetylaminofluorene (AAF) adduct located in a short, heteroduplex sequence was analyzed in Saccharomyces cerevisiae. The strains used were proficient or deficient for the activity of DNA polymerase zeta (REV3 and rev3delta, respectively) in a mismatch and nucleotide excision repair-defective background (msh2delta rad10delta). The plasmid design enabled the determination of the frequency with which translesion synthesis (TLS) and mechanisms avoiding the adduct by using the undamaged, complementary strand (damage avoidance mechanisms) are invoked to complete replication. To this end, a hybridization technique was implemented to probe plasmid DNA isolated from individual yeast transformants by using short, 32P-end-labeled oligonucleotides specific to each strand of the heteroduplex. In both the REV3 and rev3delta strains, the two strands of an unmodified heteroduplex plasmid were replicated in approximately 80% of the transformants, with the remaining 20% having possibly undergone prereplicative MSH2-independent mismatch repair. However, in the presence of the AAF adduct, TLS occurred in only 8% of the REV3 transformants, among which 97% was mostly error free and only 3% resulted in a mutation. All TLS observed in the REV3 strain was abolished in the rev3delta mutant, providing for the first time in vivo biochemical evidence of a requirement for the Rev3 protein in TLS.


Interference screws should be shorter than the hamstring tendon graft in the bone tunnel for best fixation.
Interference screw fixation of hamstring tendon grafts in bone has to overcome the challenges that tendons have a slippery surface and viscoelastically adapt under pressure. As the typical failure mode of the graft is to slip past the interference screw, it was hypothesized that the position and configuration of the graft end may be of influence on the fixation strength. Different configurations of the graft ending and its effect to primary fixation with interference screws after viscoelastic adaptation were tested in six groups: I: graft and the screw inserted at the same depth, II/III: the graft overlaps the tip of the screw (interference screw of 28 and 19 mm in length, respectively), IV: strengthening of the graft ending with additional suture knots, V: Endopearl, respectively, and VI: effect of partial retraction of the screw after excessive insertion. In vitro tests were performed with fresh calf tendon grafts and interference screws in bone tunnels (fresh porcine distal femur) all of 8 mm in diameter. The relative position of the graft ending to the tip of the interference screw thereby was recognized as a significant factor on pullout forces. Further strengthening at the graft endings with additional suture knots or an Endopearl device could improve primary hold as well. Better fixation strength is achieved if the tip of interference screw does not extend past the end of a tendon graft. Enforcement of the tendon end with sutures or an implant can further improve fixation.


What is the appropriate approach to treating women with incurable cervical cancer?
Women should not die of cervical cancer...but they do. Most cases of cervical cancer are preventable and, if caught early, highly curable. Despite this, cervical cancer is the second most common cause of cancer death in women worldwide. Unfortunately, cure is less likely when the disease is diagnosed at an advanced stage. Treatment of locally advanced disease often has serious lifelong side effects, including cystitis, proctitis, intestinal strictures, fistulas, vaginal agglutination/sexual dysfunction, and chronic pain. Unresectable recurrent disease may be associated with all these symptoms, in addition to disturbing malodorous discharge and bleeding and a life expectancy of less than 6 to 10 months. Supportive/palliative care interventions are needed at diagnosis and recurrence. Honest, conflict-free conversations about potential for response and expectations are needed for women with recurrent disease to help preserve and improve quality of life and avoid treatments that offer no benefit. Aims of trial design should include not only molecular targeting but also supportive care objectives, such as reducing pain, anxiety, depression, cachexia, and fatigue. A β-adrenergic blockade could potentially be part of these intervention trials. This article addresses the following questions and issues: whether therapeutic treatment of incurable cervical cancer is ever appropriate, the common symptoms of recurrent cervical cancer, the quantifying benefits of treatment, decision aids in treatment planning, doctor-patient candor, and integrating psychosocial factors into treatment.


Rhesus monkey rhadinovirus ORF57 induces gH and gL glycoprotein expression through posttranscriptional accumulation of target mRNAs.
Open reading frame 57 (ORF57) of gamma-2 herpesviruses is a key regulator of viral gene expression. It has been reported to enhance the expression of viral genes by transcriptional, posttranscriptional, or translational activation mechanisms. Previously we have shown that the expression of gH and gL of rhesus monkey rhadinovirus (RRV), a close relative of the human Kaposi's sarcoma-associated herpesvirus (KSHV), could be dramatically rescued by codon optimization as well as by ORF57 coexpression (J. P. Bilello, J. S. Morgan, and R. C. Desrosiers, J. Virol. 82:7231-7237, 2008). We show here that ORF57 coexpression and codon optimization had similar effects, except that the rescue of expression by codon optimization was temporally delayed relative to that of ORF57 coexpression. The transfection of gL mRNA directly into cells with or without ORF57 coexpression and with or without codon optimization recapitulated the effects of these modes of induction on transfected DNA. These findings suggested an important role for the enhancement of mRNA stability and/or the translation of mRNA for these very different modes of induced expression. This conclusion was confirmed by several different measures of gH and gL mRNA stability and accumulation with or without ORF57 coexpression and with or without codon optimization. Our results indicate that RRV gH and gL expression is severely limited by the stability of the mRNA and that ORF57 coexpression and codon optimization independently induce gH and gL expression principally by allowing accumulation and translation of these mRNAs.


Targeting the Peroxisome Proliferator-Activated Receptor-γ to Counter the Inflammatory Milieu in Obesity.
Adipose tissue, which was once viewed as a simple organ for storage of triglycerides, is now considered an important endocrine organ. Abnormal adipose tissue mass is associated with defects in endocrine and metabolic functions which are the underlying causes of the metabolic syndrome. Many adipokines, hormones secreted by adipose tissue, regulate cells from the immune system. Interestingly, most of these adipokines are proinflammatory mediators, which increase dramatically in the obese state and are believed to be involved in the pathogenesis of insulin resistance. Drugs that target peroxisome proliferator-activated receptor-γ have been shown to possess anti-inflammatory effects in animal models of diabetes. These findings, and the link between inflammation and the metabolic syndrome, will be reviewed here.


The potential protective effect of two actinomycete extracts against carbon tetrachloride-induced hepatotoxicity in rats.
The aim of this study was to investigate the potential protective effect of two extracts derived from two soil actinomycete strains, designated S19 and G30, against CCl4-induced hepatotoxicity in male rats. Sixty-four male rats were divided into four groups of 16 rats per group. The first group was a control group given corn oil and the nutritive medium which is composed of a mixture of the two used media. The second group received CCl4 only, the third group was administered CCl4 and the extract S19, and the fourth group was administered CCl4 and the extract G30. The results were taken after a treatment period of 8 weeks. Our data demonstrated that the two actinomycete extracts significantly (P < 0.01) lowered the CCl4-induced elevation of serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) after 8 weeks of treatment. The extract S19 had no effect on serum lactate dehydrogenase (LDH) and total bilirubin, whereas the extract G30 significantly decreased (P < 0.01) the elevated levels of these parameters in the serum, especially after 4 weeks of treatment. The levels of hepatic glutathione (GSH), glutathione peroxidase (GSH-Px), peroxidase (Px), catalase (CAT), and superoxide dismutase (SOD) significantly increased (P < 0.01), while those of malondialdehyde (MDA) markedly decreased in rats treated with the two extracts. Furthermore, histopathological lesions in the liver, including necrosis, inflammatory cell infiltration, hydropic degeneration, and congestion of the central vein, were partially reversed by treatment with the two microbial extracts. Our results provided evidence for the protective effect of the two used actinomycete extracts against CCl4-induced liver damage occurred through the reduction of oxidative stress and improvement of antioxidant defense markers.


Multicentre evaluation of an emergency department asthma care pathway for adults.
We sought to determine whether a standardized emergency department (ED) asthma care pathway (ACP) for adults would be accepted by ED staff, improve adherence to Canadian ED asthma management guidelines and improve patient outcomes. Ten Ontario hospital EDs (5 intervention, 5 control) participated in a 5-month pre-post intervention study. Emergency department management, admissions, repeat ED visits and ED length of stay were compared between sites and by ACP use versus nonuse at intervention sites. The ACP was used in 101 of 383 visits (26.4%) at 5 intervention sites. Use of the ACP varied significantly between sites, ranging from 6% to 60% (p < 0.001). When compared with control sites, there were significant increases in the use of metered dose inhalers (MDIs), inhaled steroids, referrals, documentation of teaching, patient recollection of teaching (all with a p < 0.001) and oxygen (p = 0.001). Use of peak expiratory flow rate (PEFR) measurements decreased in both intervention and control sites. Increased PEFR documentation and systemic steroid use in the ED and on discharge were only found in patients who were on the ACP at intervention sites. Admissions increased from 3.9% to 9.4% at intervention sites in contrast to control sites, where they remained fairly stable (p = 0.016), but did not differ by ACP use. The length of stay for discharged patients increased by a mean of 16 minutes for ACP patients at intervention sites (p = 0.002). There were no statistically significant differences in repeat ED visits. Adoption of a standardized ED ACP for adults is highly variable. Despite modest uptake, which averaged 26%, beneficial changes in specific aspects of asthma care delivery were found, notably in referrals and recollection of teaching done during the ED visit, without a substantial increase in ED length of stay. These changes may lead to improvements in outcomes, such as reduced relapse rates, which this study was not designed or powered to detect. Provincial and national implementation strategies that address barriers to clinical pathway adoption are warranted and have the potential to improve adherence to guidelines and outcomes for asthma patients.


A unique tubulin antibody which disrupts particle movement in squid axoplasm.
Microtubules have been demonstrated to be a substrate for organelle transport and particle translocation in vitro and in vivo. Subsequent to a previous report of inhibition of axonal transport of exogenous tracers in vivo using antiserum NS-20 against tubulin (Johnston et al: Brain Res. 1986), we now show disruption of particle movement in extruded squid axoplasm using this unique immunological probe. Using video-enhanced contract-differential interference contrast (AVEC-DIC) microscopy, we examined the properties of particle movement along microtubules and demonstrated that both the velocity of particle movement and the numbers of particles moving are decreased in the presence of NS-20 antiserum or NS-20 affinity-purified antibodies but not in the presence of another antiserum against tubulin. The amount of microtubule substrate does not change in the presence of any of the antisera. In conclusion, we suggest that NS-20 antibodies bind near or at a site on the tubulin molecule which is critical in the mechanism of particle transport, and provide a direct immunological probe to examine the mechanism of microtubule involvement in axonal transport.


Harlequin baby with ecthyma gangrenosum.
Pseudomonas aeruginosa bacteremia or sepsis often occurs in hospitals, affecting mainly children with underlying disease. Ecthyma gangrenosum is classically considered a pathognomonic sign of sepsis by P. aeruginosa. The harlequin baby, a severe variant of ichthyosis, occurs rarely, and these infants are at high risk of cutaneous infections and sepsis. We herein report a harlequin baby who developed ecthyma gangrenosum.


Improving Engagement Among Patients With Chronic Cardiometabolic Conditions Using mHealth: Critical Review of Reviews.
The burden imposed by cardiometabolic diseases remains a principal health care system concern. Integration of mobile health (mHealth) interventions is helpful for telemonitoring of these patients, which enables patients to be more active and take part in their treatment, while being more conscious and gaining more control over the outcomes. However, little is known about the degree to which users engage, and the extent to which this interaction matches the usage pattern for which mHealth interventions were designed. The aim of this study was to describe the characteristics and results of studies on mHealth solutions that measured the effects of interventions with patient engagement in the context of chronic cardiometabolic diseases. A critical review of systematic reviews was conducted to recover data on interventions focused on the engagement of patients with chronic cardiometabolic diseases using mHealth technologies. Articles (from January 1, 2010) were searched in the Medlars Online International Literature Medline (Medline/Pubmed), Embase, Cochrane Library, PsycINFO, and Scielo databases. Only studies that quantified a measure of engagement by patients with cardiometabolic disease were included for analysis. The Critical Appraisal Skills Programme (CASP) was used to determine included studies considering the quality of the data provided. The Scottish Intercollegiate Guidelines Network (SIGN) checklist was used to assess the quality of the evidence according to the methodology used in the studies reviewed. Engagement was defined as the level of patient implication or participation in self-care interventions. Engagement measures included number of logs to the website or platform, frequency of usage, number of messages exchanged, and number of tasks completed. Initially, 638 papers were retrieved after applying the inclusion and exclusion criteria. Finally, only three systematic reviews measuring engagement were included in the analysis. No reviews applying a meta-analysis approach were found. The three review articles described the results of 10 clinical trials and feasibility studies that quantified engagement and met the inclusion criteria assessed through CASP. The sample size varied between 6 and 270 individuals, who were predominantly men. Cardiac disease was the principal target in the comparison of traditional and mHealth interventions for engagement improvement. The level of patient engagement with mHealth technologies varied between 50% and 97%, and technologies incorporating smartphones with a reminder function resulted in the highest level of engagement. mHealth interventions are an effective solution for improving engagement of patients with chronic cardiometabolic diseases. However, there is a need for advanced analysis and higher-quality studies focused on long-term engagement with specific interventions. The use of smartphones with a single app that includes a reminder function appears to result in better improvement in active participation, leading to higher engagement among patients with cardiometabolic diseases.


Psychiatric aspects of Hansen's disease (leprosy).
Hansen's disease or leprosy is a major worldwide public health problem that has profound psychological effects on its victims. This paper is the first systematic study of psychiatric patients with Hansen's disease in 30 years and the first to use current diagnostic criteria. Because the incidence of Hansen's disease appears to be rising, the implications of psychiatric diagnosis in Hansen's disease patients and treatment considerations for the future are discussed.


The contributions of the actin machinery to endocytic membrane bending and vesicle formation.
Branched and cross-linked actin networks mediate cellular processes that move and shape membranes. To understand how actin contributes during the different stages of endocytic membrane reshaping, we analyzed deletion mutants of yeast actin network components using a hybrid imaging approach that combines live imaging with correlative microscopy. We could thus temporally dissect the effects of different actin network perturbations, revealing distinct stages of actin-based membrane reshaping. Our data show that initiation of membrane bending requires the actin network to be physically linked to the plasma membrane and to be optimally cross-linked. Once initiated, the membrane invagination process is driven by nucleation and polymerization of new actin filaments, independent of the degree of cross-linking and unaffected by a surplus of actin network components. A key transition occurs 2 s before scission, when the filament nucleation rate drops. From that time point on, invagination growth and vesicle scission are driven by an expansion of the actin network without a proportional increase of net actin amounts. The expansion is sensitive to the amount of filamentous actin and its cross-linking. Our results suggest that the mechanism by which actin reshapes the membrane changes during the progress of endocytosis, possibly adapting to varying force requirements.


Should leptin replace insulin as a lifetime monotherapy for diabetes type 1 and 2?
Evidence accumulated during the last decade has affirmed that adipocyte leptin insufficiency in the hypothalamus is the primary etiological factor in the pathogenesis of diabetes type 1 and 2 and related metabolic morbidities. Leptin insufficiency disrupts the relay of hypothalamic regulatory information along three descending pathways to the organs in the periphery that normally participate in maintenance of glucose homeostasis on a minute-to-minute basis throughout lifetime. Reinstatement of leptin sufficiency in the hypothalamus by either systemic or central injections, or its provision selectively in the hypothalamus with the aid of gene therapy extinguished hyperglycemia and normalized blood glucose stably during the entire course of treatment in a variety of animal models of diabetes type 1 and 2. In follow-up clinical trials, twice daily leptin treatment in leptinopenic and insulinopenic type 1 diabetics and leptinopenic and hyperinsulinemic type 2 diabetics with congenital lipodystrophy or acquired lipoatrophy normalized blood glucose without any discernible adverse effects during the extended course of treatment. Taken together, these findings have amply endorsed the efficacy of leptin therapy to restore glucose homeostasis in insulin-deficient as well as hyperinsulinemic diabetic patients. Consequently, restoration of optimal hypothalmic signaling to reinstate glucose homeostasis with leptin is a highly suitable new therapeutic strategy to ameliorate diabetes type 1 and 2 for the lifetime and to replace the currently in vogue insulin monotherapy. In view of the relentless challenges posed by the worldwide epidemic of diabetes and soaring treatment costs, taken together with the well-known shortcomings of therapies based on restoring insulin signaling, it is highly critical and timely to undertake new clinical trials that ascertain appropriate dosage and route of leptin delivery to the hypothalamus capable of safely sustaining stable glycemia for lifetime.


A role for mammalian Ubc6 homologues in ER-associated protein degradation.
Integral membrane and secretory proteins which fail to fold productively are retained in the endoplasmic reticulum and targeted for degradation by cytoplasmic proteasomes. Genetic and biochemical analyses suggest that substrates of this pathway must be dislocated across the membrane of the endoplasmic reticulum (ER) by a process requiring a functional Sec61 complex and multiubiquitinylation. In yeast, the tail-anchored ubiquitin-conjugating enzyme Ubc6p, which is localized to the cytoplasmic surface of the ER, participates in ER-associated degradation (ERAD) of misfolded proteins. Here we describe the identification of two families of mammalian Ubc6p-related proteins. Members of both families are also located in the ER membrane and display a similar membrane topology as the yeast enzyme. Furthermore we show that expression of elevated levels of wild-type and dominant-negative alleles of these components affects specifically ERAD of the alpha subunit of the T-cell receptor and a mutant form of the CFTR protein. Similarly, we describe that the expression level of Ubc6p in yeast is also critical for ERAD, suggesting that the Ubc6p function is highly conserved from yeast to mammals.


Paediatric airway stenosis: laryngotracheal reconstruction or cricotracheal resection?
Modern surgical management of paediatric laryngotracheal stenosis includes a wide variety of surgical procedures. These can broadly be divided into two groups. First, laryngotracheal reconstruction (LTR) procedures in which the cricoid cartilage is split and the framework is expanded with various combinations of cartilage grafts and stents; and second, cricotracheal resection (CTR) where a segmental excision of the stenotic segment is done and an end-to-end anastomosis is performed. In this article we review the literature and our experience and discuss the relative indications for CTR and LTR in children. High decannulation rates have been reported for CTR; however, it remains a more extensive procedure than LTR involving extensive tracheal mobilization. If the tracheostomy site is included in the resection then a significant length of trachea is excised. Alternatively, LTR with cartilage grafting can precisely correct a specific stenosis with minimum morbidity and high decannulation rates for grade 2 and selected grade 3 stenosis. For the more severe stenosis treatment with LTR has been less successful. Retrospective data from this institution suggests that the children with grade 4 stenosis treated with LTR are more likely to require a subsequent open procedure to achieve decannulation than those treated with CTR. LTR is a less extensive procedure and is preferred for grade 2, selected grade 3 stenosis. CTR is the preferred option for grade 4 and severe grade 3 stenosis with a clear margin between the stenosis and the vocal cords.


Calcium calmodulin kinase II activity is required for cartilage homeostasis in osteoarthritis.
WNT ligands can activate several signalling cascades of pivotal importance during development and regenerative processes. Their de-regulation has been associated with the onset of different diseases. Here we investigated the role of the WNT/Calcium Calmodulin Kinase II (CaMKII) pathway in osteoarthritis. We identified Heme Oxygenase I (HMOX1) and Sox-9 as specific markers of the WNT/CaMKII signalling in articular chondrocytes through a microarray analysis. We showed that the expression of the activated form of CaMKII, phospho-CaMKII, was increased in human and murine osteoarthritis and the expression of HMOX1 was accordingly reduced, demonstrating the activation of the pathway during disease progression. To elucidate its function, we administered the CaMKII inhibitor KN93 to mice in which osteoarthritis was induced by resection of the anterior horn of the medial meniscus and of the medial collateral ligament in the knee joint. Pharmacological blockade of CaMKII exacerbated cartilage damage and bone remodelling. Finally, we showed that CaMKII inhibition in articular chondrocytes upregulated the expression of matrix remodelling enzymes alone and in combination with Interleukin 1. These results suggest an important homeostatic role of the WNT/CaMKII signalling in osteoarthritis which could be exploited in the future for therapeutic purposes.


Failed appointments. Who misses them, why they are missed, and what can be done.
Failure to keep appointments is disruptive to the health care delivery system in several ways. Rates of missed appointments in a variety of practice settings demonstrate a significant problem. The patient most likely to fail to keep a given appointment is one who is young, comes from a low socioeconomic group, has a large, unstable family, and has previously broken appointments. In addition, this patient will most likely have no significant ongoing relationship with a single physician, may have been referred from an emergency room, will have been scheduled for his or her appointment at a distant time, may well have forgotten about the appointment or thought it was scheduled for another time, and will feel little sense of urgency about keeping the appointment. Several effective methods to reduce appointment-breaking behavior are discussed. Mailed appointment reminders are both effective and cost-efficient. Improved communication between patient and physician combined with personal interest and attention an produce a positive effect on the appointment-keeping behavior of a patient. Finally, the pragmatic details of appointment scheduling can be altered to reduce the disruptive effect of the missed appointment by such methods as predictive overbooking based on individual patient characteristics, use of the modified wave scheduling technique, and elimination of the automatic reappointing of patients who have previously broken appointments.


Carcinogen exposure differentially modulates RAR-beta promoter hypermethylation, an early and frequent event in mouse lung carcinogenesis.
The retinoic acid receptor beta (RAR-beta) gene encodes one of the primary receptors for retinoic acid, an important signaling molecule in lung growth, differentiation and carcinogenesis. RAR-beta has been shown to be down-regulated by methylation in human lung cancer. We have used previously lung tumors induced in mice to evaluate the timing and effect of specific carcinogen exposures on targeting genes altered in human lung cancer. These studies were extended to characterize the role of methylation of the RAR-beta gene in murine lung cancers. After treatment with the demethylating agent 5-aza-2'-deoxycytidine (DAC), RAR-beta was re-expressed in silenced cell lines or expressed at a higher rate than without DAC, supporting methylation as the inactivating mechanism. Bisulfite sequencing detected dense methylation in the area of the CpG island that contained the 5' untranslated region and the first translated exon in non-expressing cell lines, compared with minimal and heterogeneous methylation in normal mouse lung. Methylation-specific PCR revealed that this gene is targeted differentially by carcinogen exposures with the detection of methylated alleles in virtually all primary tumors associated with cigarette smoke or 4-methylnitrosamino-1-(3-pyridyl)-butanone (NNK) in contrast to half of tumors induced by methylene chloride or vinyl carbamate. RAR-beta methylation was also detected in 54% of preneoplastic hyperplasias induced by treatment with NNK. Bisulfite sequencing of both premalignant and malignant lesions detected dense methylation in the same area observed in cell lines, substantiating that this gene is functionally inactivated at the earliest histologic stage of adenocarcinoma development. These studies demonstrate that aberrant methylation of RAR-beta is an early and common alteration in murine lung tumors induced by several environmentally relevant exposures.


Chaplains and telechaplaincy: best practices, strengths, weaknesses-a national study.
Telechaplaincy is the use of telecommunications and virtual technology to deliver religious/spiritual care. It has been used for decades, but chaplains' understanding of telehealth lags behind other disciplines. The purpose of this study was to describe the use of telechaplaincy in the United States and chaplains' perceptions of the practice. Researchers surveyed chaplains through chaplain-certifying-body email-listservs, then conducted in-depth interviews with 36 participants identified through maximum variation sampling. Quantitative analysis and qualitative, thematic analysis were conducted. Quantitative results show that in 2019, approximately half of surveyed chaplains performed telechaplaincy. Rural chaplains were more likely to have practiced. Chaplains who had not practiced were more willing to try if they believed it was effective at meeting religious/spiritual needs. Qualitative findings describe chaplains' perceptions of strengths, weaknesses, and best practices.


Predicting the outcomes of shunt implantation in patients with post-traumatic hydrocephalus and severe conscious disturbance: a scoring system based on clinical characteristics.
Post-traumatic hydrocephalus is a common complication secondary to traumatic brain injury. It can cause cerebral metabolic impairment and dysfunction. Therefore, timely treatment with shunt implantation is necessary. However, the outcomes of shunt surgery in patients with post-traumatic hydrocephalus combined with disturbance of consciousness are doubtful. The objective was to develop a predictive model that uses the information available before surgery to predict the outcome of shunt implantation in such patients. Retrospectively collected data were used to develop a clinical prediction model. The model was derived from 59 patients using logistic regression analysis, and then it was evaluated by the area under the receiver operating characteristic curve and Hosmer-Lemshow test. A validation cohort verified the model. Four independent predictors were identified: age < 50 years, mild hydrocephalus, Glasgow Coma Scale scores 9-12 at the time of injury, and time interval from trauma to shunting < 3 months. We calculated the total score and defined the patients into three groups: low-probability (0-10 points), medium-probability (11-16 points), and high-probability (17-30 points). The rates of improved outcomes in the three groups were 14.3%, 52.6%, and 94.7%, respectively (P < 0.0001). The correlative rates of the validation cohort were 21.4%, 54.5%, and 85.7%. The prognostic model showed good discrimination (area under the receiver operating characteristic curve = 0.869) and calibration (Hosmer-Lemshow test, P = 0.391). The developed predictive model can identify patients with post-traumatic hydrocephalus combined with disturbance of consciousness who can benefit from shunt implantation. Therefore, our prognostic model can predict the outcomes of patients with post-traumatic hydrocephalus and disturbance of consciousness after shunt surgery.


Resting state activity and the "stream of consciousness" in schizophrenia--neurophenomenal hypotheses.
Schizophrenia is a multifaceted disorder with various symptoms including auditory hallucinations, egodisturbances, passivity phenomena, and delusions. Recent neurobiological approaches have focused on, especially, the abnormal contents of consciousness, the "substantive parts" as James said, to associate them with the neural mechanisms related to sensory, motor, and cognitive functions, and the brain's underlying stimulus-induced or task-evoked activity. This leaves open, however, the neural mechanisms that provide the temporal linkage or glue between the various contents, the transitive parts that makes possible the "stream of consciousness." Interestingly, schizophrenic patients seem to suffer from abnormalities specifically in the "transitive parts" when they experience contents as temporally disconnected or fragmented which in phenomenological psychiatry has been described as "temporal fragmentation." The aim of this article is to develop so-called neurophenomenal hypothesis about the direct relationship between phenomenal features of the "stream of consciousness," the "transitive parts," and the specific neuronal mechanisms in schizophrenia as based on healthy subjects. Rather than emphasizing stimulus-induced and task-evoked activity and sensory and lateral prefrontal cortical regions as in neurocognitive approaches with their focus on the "substantive parts," the focus shifts here to the brain's intrinsic activity, its resting state activity, which may account for the temporal linkage or glue between the contents of consciousness, the transitive parts.


Dynamic mechanical loading and growth factors influence chondrogenesis of induced pluripotent mesenchymal progenitor cells in a cartilage-mimetic hydrogel.
Human induced pluripotent stem cells (iPSCs) have emerged as a promising alternative to bone-marrow derived mesenchymal stem/stromal cells for cartilage tissue engineering. However, the effect of biochemical and mechanical cues on iPSC chondrogenesis remains understudied. This study evaluated chondrogenesis of induced pluripotent mesenchymal progenitor cells (iPS-MPs) encapsulated in a cartilage-mimetic hydrogel under different culture conditions: free swelling versus dynamic compressive loading and different growth factors (TGFβ3 and/or BMP2). Human iPSCs were differentiated into iPS-MPs and chondrogenesis was evaluated by gene expression (qPCR) and protein expression (immunohistochemistry) after three weeks. In pellet culture, both TGFβ3 and BMP2 were required to promote chondrogenesis. However, the hydrogel in growth factor-free conditions promoted chondrogenesis, but rapidly progressed to hypertrophy. Dynamic loading in growth factor-free conditions supported chondrogenesis, but delayed the transition to hypertrophy. Findings were similar with TGFβ3, BMP2, and TGFβ3 + BMP2. Dynamic loading with TGFβ3, regardless of BMP2, was the only condition that promoted a stable chondrogenic phenotype (aggrecan + collagen II) accompanied by collagen X down-regulation. Positive TGFβRI expression with load-enhanced Smad2/3 signaling and low SMAD1/5/8 signaling was observed. In summary, this study reports a promising cartilage-mimetic hydrogel for iPS-MPs that when combined with appropriate biochemical and mechanical cues induces a stable chondrogenic phenotype.


Evidence that CB-1 and CB-2 cannabinoid receptors mediate antinociception in neuropathic pain in the rat.
The roles of the two cannabinoid receptor subtypes, CB-1 and CB-2, have not been clarified in cannabinoid-mediated analgesia. We investigated the efficacy of the non-selective cannabinoid receptor agonist CP55,940 in the modulation of responses in the rat to both acute pain (tail flick) and neuropathic pain (tactile allodynia following chronic L5/6 spinal nerve ligation). Responses were also assessed in the presence of the CB-1 antagonist SR141716A (SR1) and the CB-2 antagonist SR144528 (SR2). CP55,940 attenuated tactile allodynia (ED(50) 0.04 mg/kg i.t. (95% CI 0.032-0.044 mg/kg), 0.12 mg/kg i.p. (95% CI 0.10-0.15 mg/kg)) and induced thermal antinociception (ED(50) tail flick 0.07 mg/kg i.t. (95% CI 0.05-0.10 mg/kg), 0.17 mg/kg i.p. (95% CI 0.11-0.26 mg/kg)). SR1 0.5 mg/kg i.t. attenuated the antinociceptive effect of CP55,940 in both modalities. However, SR1 1.0 mg/kg i.p. decreased tail flick latency but had no effect on tactile allodynia antinociception. In contrast, SR2 1.0 mg/kg i.p. significantly decreased the effect of i.p. CP55,940 on both tail flick antinociception and tactile allodynia (P<0.005). The combination of SR1 and SR2 (i.p.) had an additive effect in decreasing the antinociception induced by CP55,940 on tail flick responses (P<0.005). These results suggest a role for CB-2 receptor-mediated antinociception in both acute and neuropathic pain in addition to centrally located CB-1 mechanisms.


Chloroplast and mitochondrial DNA in a brown alga Egregia menziesii.
Chloroplasts and mitochondria of the brown alga Egregia menziesii were studied with the electron microscope. In both organelles, 15-25-A fibrils with DNA characteristics are found within areas of electron transparency. In each chloroplast there are two DNA-containing areas, one at each tip of the chloroplast. This localization, the shape and size of each DNA-containing area, and its close association with lamellae in a nondividing chloroplast are noted. One or occasionally two DNA-containing areas are found within the mitochondrion and they are compared with a similar structure in the chloroplast.